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1.
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2.
Our objective was to develop a lipopolysaccharide (LPS) inflammation model in calves to evaluate the acute-phase response with respect to the release of pro-inflammatory cytokines and acute-phase proteins, fever development and sickness behaviour. Fourteen 4-week-old male Holstein Friesian calves were included and randomly assigned to a negative control group (n = 3) and an LPS-challenged group (n = 11). The latter received an intravenous bolus injection of 0.5 μg of LPS/kg body weight. Blood collection and clinical scoring were performed at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 12, 18, 24, 28, 32, 48, 54 and 72 h post LPS administration (p.a.). In the LPS group, the following clinical signs were observed successively: tachypnoea (on average 18 min p.a.), decubitus (29 min p.a.), general depression (1.75 h p.a.), fever (5 h p.a.) and tachycardia (5 h p.a.). Subsequent to the recovery from respiratory distress, general depression was prominent, which deteriorated when fever increased. One animal did not survive LPS administration, whereas the other animals recovered on average within 6.1 h p.a. Moreover, the challenge significantly increased plasma concentrations of tumour necrosis factor-α, interleukin 6, serum amyloid A and haptoglobin, with peaking levels at 1, 3.5, 24 and 18 h p.a., respectively. The present LPS model was practical and reproducible, caused obvious clinical signs related to endotoxemia and a marked change in the studied inflammatory mediators, making it a suitable model to study the immunomodulatory properties of drugs in future research.  相似文献   

3.
Neutrophils are the predominant cells recruited in the airways of horses suffering from heaves. These cells have been shown to express arginase in some species. The metabolism of l-arginine is thought to be involved in chronic inflammation, and airway obstruction and remodeling. The aim of this study was to assess the expression, regulation, activity, and functional role of arginase isoforms in equine neutrophils. Arginase I, arginase II, ornithine decarboxylase (ODC) and ornithine aminotransferase (OAT) expression were assessed in resting and stimulated (IL-4, LPS/fMLP, PMA; 5 and 18 h) blood neutrophils using quantitative PCR. Arginase expression was also studied by Western blot and enzyme activity assay. The effect of nor-NOHA (1 mM), a specific arginase inhibitor, was assessed on arginase activity in vitro and ex vivo on neutrophil's inflammatory gene expression and viability. Results showed that equine neutrophils constitutively express arginase isoform 2, ODC and OAT. Neutrophil ex vivo stimulation did not induce arginase I or influence arginase II mRNA expression. Ex vivo inhibition of arginase activity by nor-NOHA had no effect on neutrophils inflammatory gene expression induced by LPS/fMLP (5 h) but significantly reversed the cell loss observed after this stimulation.  相似文献   

4.
Intravenous injection of microparticles (MPs) is a tool to reveal susceptibility to pulmonary hypertension (PH) syndrome (PHS, ascites) in broilers. After injection MPs get lodged in pulmonary arterioles and cause localized inflammation. To examine the expression of chemokines/cytokines during the MP-induced pulmonary inflammatory response, lungs were collected from 4-week-old broilers (6/line/time point) from the PHS-resistant (RES) and -susceptible (SUS) broilers before (0 h) and after (2, 6, 12, 24, and 48 h) MP injection and analyzed using quantitative RT-PCR. In both lines, expression of interleukin-1β (IL-1β), IL-6, IL-8, and K60 increased from 0 to 6 h, reached peak levels at 6 and 12 h, and decreased thereafter, whereas IL-4 and interferon gamma (IFN-γ) expression remained elevated past 12 h. Lungs from the RES line broilers had higher expression (P < 0.05) of IL-1β and IL-6 at 2, 6, and 12 h; higher IL-8 at 6 and 12 h; higher K60 at 6, 12, and 24 h; higher IL-4 at 12, 24, and 48 h and higher IFN-γ expression at 6 and 48 h post-MP injection than SUS line broilers. Higher expression of chemokines/cytokines in RES compared to SUS line lungs may explain the ability of RES line broilers to effectively counteract the MP-induced PH and resolve the vascular occlusion.  相似文献   

5.
This study investigated the effects of transportation stress on blood concentrations of the main pro-inflammatory cytokines (interleukins IL-1β, IL-2 and IL-6; tumour necrosis factor-α) and anti-inflammatory cytokines (IL-4 and IL-10) and the expression of these cytokines and their receptors in the thymus. Pigs were assessed after 1, 2 and 4 h of transportation (n = 5 per group), with normal housing conditions as a control (n = 4). Serum concentrations of IL-2, IL-6 and IL-10 were highest at 1 h, whereas concentrations of IL-6 and IL-10 were significantly decreased at 4 h. Expression of these three cytokines and their receptors was also significantly altered in the thymus during transportation stress. Serum IL-10 concentrations and thymus IL-10 mRNA expression were significantly correlated. The thymus may contribute towards the regulation of cytokines in pigs during transportation.  相似文献   

6.
Activated neutrophils live longer, produce toxic metabolites and cause considerable tissue injury, which is central to the pathogenesis of many inflammatory conditions. Retinoids are a class of lipophilic compounds with anti-inflammatory effects. We examined the effect of retinoic acid on apoptosis in normal and activated neutrophils. Our results showed that treatment with 1 μg/ml Escherichia coli lipopolysaccharide (LPS) for 12 and 36 h delayed the spontaneous neutrophil apoptosis compared to untreated cells. But exposure of LPS-treated cells to retinoic acid (1 and 5 μM) abolished the inhibitory effects of LPS on neutrophil apoptosis in a concentration-dependant manner based on annexin V staining, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, light and electron microscopy. These results show that retinoic acid increases apoptosis in activated canine neutrophils and this effect could enhance the resolution of inflammation in vivo.  相似文献   

7.
Trefoil factors (TFF) are small peptides produced by goblet cells, which are crucial for epithelial restitution. In humans with inflammatory bowel disease (IBD), TFF expression is up-regulated as part of an unspecific repair mechanism. The goal of this study was to assess TFF gene expression in the gastrointestinal tract from dogs with IBD compared to healthy controls. Preliminary assessment by PCR revealed TFF1 and 3 expression in the small and large intestine, whereas TFF2 was amplified only in the stomach. Subsequent RT-qPCR (with relative quantification against 3 reference genes) on endoscopic duodenal (IBD n = 22, healthy controls n = 18) and colonic (IBD n = 12, controls n = 11) biopsies revealed that TFF1 expression was significantly up-regulated in the duodenum from IBD dogs (Mann–Whitney p = 0.001), whereas TFF3 expression was significantly lower in IBD colon compared to controls (t-test p = 0.018).This study demonstrates evidence for dysregulation of TFF gene expression in canine IBD. Up-regulation of TFF1 could signify ectopic expression as a compensatory repair-mechanism, whereas down-regulation of TFF3 could contribute to defective epithelial barrier function, respectively. Whether this is a cause or consequence of IBD could not be established.  相似文献   

8.
This study was designed to determine if the activity of paraoxonase (PON1), an antioxidant enzyme that works as a negative acute phase reactant, is a better predictor for the clinical recovery of leishmaniotic dogs receiving standard treatments compared with inflammatory markers such as C reactive protein (CRP) and electrophoretic fractions. For this purpose we tested 20 healthy dogs (controls) and 39 leishmaniotic dogs classified as sick (group A, n = 23) or severely sick (group B, n = 16) and tested at admission and after 3, 7, 14, 21, 28, 35 and 42 days.At admission, CRP and electrophoresis were altered in both groups, while PON1 activity was abnormal only in group B. There were no differences related to the outcome (mortality, complications or time of recovery). PON1 activity normalized in about 2 weeks in dogs that had abnormal values at admission and a final positive outcome; CRP normalized in 4–6 weeks and electrophoretic fractions were still altered after 6 weeks. The results show that, at admission, inflammatory markers did not predict the outcome of leishmaniasis. PON1 activity decreased only in some dogs with systemic inflammation but not in those with mild leishmaniasis: when decreased, PON1 normalized earlier than other markers in dogs that responded to treatment. This finding most likely depends on the rapid decrease in oxidative phenomena. PON1 activity should therefore be tested on admission: if low values are recorded, severe inflammation may be suspected and PON1 measurement may be repeated during treatment to early identify responsive dogs.  相似文献   

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11.
Classical swine fever virus (CSFV) compromises the host immune system, causing the severe disease of pigs. Dendritic cells (DCs) are the most potent inducers of immune responses. In the present study, we investigated the functional properties of porcine monocyte-derived DCs (Mo-DCs) affected by CSFV. Results showed that the expression of surface markers of DCs such as major histocompatibility complex class II (MHC-II), CD80, CD83 and CD86 were unimpaired, but an obviously increased expression of CD172a in DCs was noticed 48 h after CSFV infection. The expression profiles of cytokines were detected in cultured Mo-DCs after various treatments for 48 h by Q-RT-PCR. The findings suggested that CSFV infection significantly increased the mRNA expression of IL-10 and TNF-α, and inhibited IL-12 expression, with little effect on IFN-α and IFN-γ expression. We further demonstrated that CSFV was incapable of activating the nuclear factor kappa B (NF-κB) in infected DCs, which was characterized by an unvaried DNA binding activity of NF-κB, the lack of translocation of p65/RelA from the cytoplasm to the nucleus and the stabilization of p65/RelA expression. Furthermore, Western blot analysis indicated that the inactivation of NF-κB was due to the failure of IκBα degradation. The data demonstrated that CSFV could be replicated in DCs and CSFV infection could modulate the secretion of crucial co-stimulatory molecules and cytokines which down-regulated maturation of DCs, without activating NF-κB in DCs. Thus, the results suggested a possible mechanism for CSFV evasion of innate host defenses, providing the basis for understanding molecular pathways in CSFV pathogenesis.  相似文献   

12.
The aim of this prospective study was to characterize the response of the coagulation system to a defined sterile localized inflammatory process.Tissue cages were implanted subcutaneously in five healthy Beagles. After 9–10 weeks, local inflammation was induced by an injection of 0.5 ml 1% carrageenan. Serial samples of tissue cage fluid (TCF) and blood were collected at 10 time points (0–168 h). Nucleated cells (NC) of TCF were counted automatically to characterize local inflammation. C-reactive protein (CRP), leukocytes and coagulation variables (PT, aPTT, fibrinogen, factor VIII, antithrombin, protein C, protein S, and d-dimers) were determined in blood samples.Carrageenan induced a significant 32-fold increase of NCs in TCF (P < 0.0001). A slight increase in leukocytes (P < 0.0001) was observed. There was a significant 1.3- to 1.5-fold increase in protein C (P = 0.0001) and protein S (P = 0.0028). CRP, secondary hemostasis and fibrinolysis did not change.The mild increase from baseline in PC/PS, may reflect a physiological counter reaction.  相似文献   

13.
The objectives of the current research were to determine the physiological effects and responses of many leukocytes following surgical castration and/(or) physical dehorning and the influence of anesthetics and analgesics in 3-month-old calves. Eighty 3-month-old Holstein bull calves were completely randomized to treatments in a 2 × 2 × 2 factorial arrangement with castration, dehorning, and anesthetic/analgesic as the main effects. Peripheral blood samples were collected just before (0) and 0.5, 1.5, 2.5, 4, 6, 24, and 72 h after the respective procedure(s) and analyzed for total leukocyte and differential counts, as well as plasma cortisol and haptoglobin concentrations. Blood from the 0, 0.5 and 24 h collections were analyzed for many ex vivo leukocyte responses. Data were analyzed using a repeated measures analysis of variance with the fixed effects of treatment, time, and the interaction of treatment × time. Pre-planned contrasts were performed to determine the effect of (1) management procedure (castration and/(or) dehorning), (2) anesthetic/analgesic, and (3) were the management procedures additive. There were treatment × time interactions (P < 0.05) on plasma cortisol and haptoglobin concentrations as well as for total leukocyte and neutrophil concentrations in blood. Castration and dehorning increased cortisol concentrations and the effect of the procedures was additive (P < 0.02). Dehorning alone elicited a greater (P < 0.05) cortisol response than castration alone. In contrast, the leukocytosis and neutrophilia was greater (P < 0.01) among castrated calves. In addition, haptoglobin concentrations at 24 h after castration were elevated (P < 0.01) in calves that were castrated. Both castration and dehorning suppressed (P = 0.04) many leukocyte responses including the secretion of tumor necrosis factor-α when whole blood cultures were stimulated with lipopolysaccharide, surface expression of l-selectin on peripheral blood neutrophils, and the oxidative burst intensity of peripheral blood neutrophils when co-cultured with an Escherichia coli. The effects of castration and dehorning on blood leukocyte counts or any of the leukocyte responses were not additive (P > 0.23). Castration and dehorning effects of plasma haptoglobin concentrations tended (P = 0.10) to be additive at 72 h after the procedure(s). Prior administration of local anesthetic and a systemic analgesic attenuated (P < 0.001) the cortisol response and prevented (P = 0.03) the observed leukocytosis, neutrophilia, and leukocyte suppression. These data suggest that calves should be castrated and dehorned on the same day rather than spreading them out across two days and calves should be administered pain relief prior to performing either procedure.  相似文献   

14.
Paraoxonase 1 (PON1) is a serum enzyme synthesised and secreted primarily by the liver. It possesses anti-inflammatory properties limiting the production of pro-inflammatory mediators. The objectives of this study were to validate three spectrophotometric assays for the quantification of PON1 activity in pig serum, and to determine if PON1 activity in porcine behaves as a negative acute phase protein (APP), decreasing in inflammatory conditions. An analytical validation using three different substrates – 5-thiobutil butyrolactone (TBBL), phenylacetate (PA) and 4-(p)-nitrophenyl acetate (pNA) – was performed. In addition, inflammation was experimentally induced in five pigs by subcutaneous injection of turpentine oil, while five control pigs were left untreated. The treated pigs showed significant increases in CRP and decreases in albumin, indicating an inflammatory condition. The three substrates used would be suitable for PON1 activity measurements in serum samples, since they offer adequate precision (coefficients of variation < 10%), sensitivity (0.01, 0.15, 0.02 U/mL for TBBL, pNA and PA respectively) and accuracy (r = 0.99). In addition, PON1 behaves as a negative APP in pigs since a significant decrease (P < 0.05) in its activity after 72 h of the induction of the inflammation was observed with all substrates.  相似文献   

15.
Non-steroidal anti-inflammatory drugs (NSAID) are a family of chemicals that function to reduce pain, fever, and inflammation, and they are commonly used in people and animals for this purpose. Currently there are no NSAIDs approved for the management of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess efficacy. A previous in vitro study examining biomarkers of inflammation identified fourteen genes that were significantly altered in response to Escherichia coli lipopolysaccharide (LPS)-induced inflammation. In the present study, five of those fourteen genes were tested in vivo to determine if the same effects observed in vitro were also observed in vivo. Plasma levels of prostaglandin E(2) (PGE(2)), an essential mediator of fever and inflammation, were also determined. Two groups of swine were stimulated with LPS with the second group also treated with flunixin meglumine. Blood was collected at 0, 1, 3, 6, 8, 24, and 48h post LPS-stimulation. The RNA was extracted from the blood and quantitative real-time-PCR (qRT-PCR) was utilized to determine the expression patterns of CD1, CD4, serum amyloid A2 (SAA2), Caspase 1, and monocyte chemoattractant protein 1 (MCP-1). The LPS-stimulated animals demonstrated a statistically significant alteration in expression of SAA2 and CD1 at 3h post-stimulation. Flunixin meglumine treated animals' demonstrated reduced expression of CD1 in comparison to the LPS-stimulated swine at 24 and 48h post LPS-stimulation. Flunixin meglumine treated animals exhibited reduced expression of SAA2 at 48h post-stimulation compared to LPS-stimulated swine. Swine treated with LPS demonstrated statistically significant increases in plasma PGE(2) at 1h post-stimulation. Swine treated with flunixin meglumine had no increase in plasma PGE(2) levels at any time. These results demonstrate that PGE(2) production, along with two out of five genes (SAA2 and CD1) have the potential to serve as early biomarkers of inflammation as well as indicators of NSAID efficacy.  相似文献   

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17.
Athletic performance is both a stress factor and an adaptive response to exercise that may be modulated by training, reduce inflammation and help prevent disease. Studies on the endocrinology of exercise and training have demonstrated the existence of an integrated metabolic network of hormone and cytokine regulation. Subsequent molecular studies have shown that repeated bouts of exercise may establish new basal levels of gene expression at rest.The Thoroughbred horse may be a useful ‘exercise model’ for inter-individual comparisons between subjects with homogeneous genetic and environmental backgrounds and similar exercise management practices. In this study, the effects of training and acute effort on gene expression were evaluated with a real time PCR approach in athletic (n = 10) and sedentary horses (n = 9), using a previously characterised panel of genes known to be highly modulated during effort (CXCL2, TLR4, IL1β, IL8, IL1RII, IL18, IL6 and CEBPβ). A ‘rest comparison’ was performed to evaluate a training effect in both groups while a ‘race comparison’ was performed in athletic horses only (before, immediately after, and 12 h after racing) to determine the effect of acute effort.The results indicated that many of the investigated genes (TLR4, IL1β, IL1RII, IL18, IL6 and CEBPβ) were expressed to a greater extent in athletic horses compared to sedentary animals when both were at rest. However, a time-course comparison in the athletic horses revealed that genes exhibiting the highest levels of expression at rest did not show significant changes after the race. The findings suggested that training may exert a conditioning on gene expression at rest leading to a more prompt response to exercise-induced stress in Thoroughbreds.  相似文献   

18.
A PCR based method was developed for the identification of ceftiofur resistance genes (blaCMY-2, blaTEM-1, and ampC) in swine bacterial pathogens. Using this method, the ceftiofur resistant (n = 76) and susceptible (n = 45) strains of Bordetella bronchiseptica, Salmonella spp., Escherichia coli, and Pasteurella multocida were screened for the presence of these three genes. The resistant genes were detected in 70% (blaTEM-1), 68% (blaCMY-2) and 45% (ampC) of the resistant isolates and in 18% (blaTEM-1), 27% (blaCMY-2), and 36% (ampC) of the susceptible isolates. Results obtained in the present study showed widespread distribution of these three resistance genes in ceftiofur-resistant swine pathogens. It was also observed that more pathogens are acquiring these resistance genes.  相似文献   

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The aim of the present work was to determine the presence of human and porcine trichinellosis in an area of Argentina historically regarded as Trichinella-free. Human blood donors (n = 216) and swine destined for consumption (n = 57) were evaluated by serological techniques (ELISA, immunofluorescence, and/or Western Blot). Muscle tissues from 26 of the pigs were evaluated for the presence of Trichinella larvae by the artificial digestion method. A questionnaire was used to collect and evaluate data on eating habits of the human population under study and on swine-raising conditions. The survey showed that 98.1% of the individuals (n = 212) were regular consumers of pork in the form of stuffed products such as sausages produced by local butchers. The seroprevalence (positive sera by at least two of the three methods) was 8.3% (n = 18) for human trichinellosis and 24.5% (n = 14) for porcine trichinellosis. Trichinella spiralis larvae were found in 2 of the 26 pigs (7.7%) with parasite loads of 0.33 and 2.4 muscle larvae per gram. Twelve swine found positive by serological and/or parasitological tests were raised under poor sanitary conditions (presence of rubbish in the surroundings, with cannibalism and scavenging behaviors, presence of rodents, etc.). Our study confirms the existence of porcine trichinellosis in an area regarded as Trichinella-free, provides supporting serological evidence of human infection in this area, and indicates that failure to report cases of trichinellosis based on inadequate surveillance can result in incorrect prevalence classification of an area.  相似文献   

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